Turnover of soil organic matter and of microbial biomass under C3-C4 vegetation change: Consideration of C fractionation and preferential substrate utilization

Two processes contribute to changes of the δ¹³C signature in soil pools: ¹³C fractionation per se and preferential microbial utilization of various substrates with different δ¹³C signature. These two processes were disentangled by simultaneously tracking δ¹³C in three pools - soil organic matter (SOM), microbial biomass, dissolved organic carbon (DOC) - and in CO₂ efflux during incubation of 1) soil after C₃-C₄ vegetation change, and 2) the reference C₃ soil.The study was done on the Ap horizon of a loamy Gleyic Cambisol developed under C₃ vegetation. Miscanthus giganteus - a perennial C₄ plant - was grown for 12 years, and the δ¹³C signature was used to distinguish between ‘old’ SOM (>12 years) and ‘recent’ Miscanthus-derived C (<12 years). The differences in δ¹³C signature of the three C pools and of CO₂ in the reference C₃ soil were less than 1‰, and only δ¹³C of microbial biomass was significantly different compared to other pools. Nontheless, the neglecting of isotopic fractionation can cause up to 10% of errors in calculations. In contrast to the reference soil, the δ¹³C of all pools in the soil after C₃-C₄ vegetation change was significantly different. Old C contributed only 20% to the microbial biomass but 60% to CO₂. This indicates that most of the old C was decomposed by microorganisms catabolically, without being utilized for growth. Based on δ¹³C changes in DOC, CO₂ and microbial biomass during 54 days of incubation in Miscanthus and reference soils, we concluded that the main process contributing to changes of the δ¹³C signature in soil pools was preferential utilization of recent versus old C (causing an up to 9.1‰ shift in δ¹³C values) and not ¹³C fractionation per se.Based on the δ¹³C changes in SOM, we showed that the estimated turnover time of old SOM increased by two years per year in 9 years after the vegetation change. The relative increase in the turnover rate of recent microbial C was 3 times faster than that of old C indicating preferential utilization of available recent C versus the old C.Combining long-term field observations with soil incubation reveals that the turnover time of C in microbial biomass was 200 times faster than in total SOM. Our study clearly showed that estimating the residence time of easily degradable microbial compounds and biomarkers should be done at time scales reflecting microbial turnover times (days) and not those of bulk SOM turnover (years and decades). This is necessary because the absence of C reutilization is a prerequisite for correct estimation of SOM turnover. We conclude that comparing the δ¹³C signature of linked pools helps calculate the relative turnover of old and recent pools.     

Origin of positive δC of emitted CO2 from soils after application of biogas residues

Bioenergy production from renewable organic material is known to be a clean energy source and therefore its use is currently much promoted in many countries. Biogas by-products also called biogas residues (BGR) are rich in partially stable organic carbon and can be used as an organic fertilizer for crop production. However so far, many environmental issues relevant when BGR are applied to agricultural land (soil C sequestration, increased denitrification and nutrient leaching) still have to be studied. Therefore a field experiment was set up to investigate the degradation of BGR and its impact on the decomposition of native soil organic matter based on a natural abundance stable isotope approach. Maize, a C₄ plant has been used as bioenergy crop, therefore the δ¹³C of total C in BGR was −16.0‰_PDB and soil organic matter was mostly derived from C₃ plant based detritus, SOM thus showed a δ¹³C of −28.4‰_PDB. Immediately after BGR application, soil-emitted CO₂ showed unexpectedly high δ¹³C of up to +23.6‰_PDB, which has never been reported earlier. A subsequent laboratory scale experiment confirmed the positive δ¹³C of soil-emitted CO₂ after BGR addition and showed that obviously, the added BGR led to a consumption of dissolved inorganic C in soils. Additionally, it was observed that the δ¹³C of CO₂ driven from inorganic C of BGR (BGR-IC) by acid treatment was +35.6‰_PDB. Therefore, we suggest that also under field conditions the transformation of BGR-IC into CO₂ contributed largely to CO₂ emissions in addition to the decomposition of organic matter, which affected both the amount and the carbon isotope signature of emitted CO₂ in the initial period after BGR application. Positive δ¹³C of inorganic C contained in BGR was attributed to processes with strong fractionation of C isotopes during anaerobic fermentation in the biogas formation process.     

C signature of CO2 evolved from incubated maize residues and maize-derived sheep faeces

Analyses of the spatial and temporal variations in the natural abundance of ¹³C are frequently employed to study transformations of plant residues and soil organic matter turnover on sites where long continued vegetation with the C₃-type photosynthesis pathway has been replaced with a C₄-type vegetation (or vice versa). One controversial issue associated with such analyses is the significance of isotopic fractionation during the microbial turnovers of C in complex substrates. To evaluate this issue, C₃-soil and quartz sand were amended with maize residues and with faeces from sheep feed exclusively on maize silage. The samples were incubated at 15 °C for 117 days (maize residues) or 224 days (sheep faeces). CO₂ evolved during incubation was trapped in NaOH and analysed for C isotopic contents. At the end of incubation, 63 and 50% of the maize C was evolved as CO₂ in the soil and sand, respectively, while 32% of the faeces C incubated with soil and with sand was recovered as CO₂. Maize and faeces showed a similar decomposition pattern but maize decomposed twice as fast as faeces. The δ¹³C of faeces was 0.3‰ lower than that of the maize residue (δ¹³C −13.4‰), while the δ¹³C of the C₃-soil used for incubation was −31.6‰. The δ¹³C value of the CO₂ recovered from unamended C₃-soil was similar or slightly lower (up to −1.5‰) than that of the C₃-soil itself except for an initial flush of ¹³C enriched CO₂. The δ¹³C values of the CO₂ from sand-based incubations typically ranged −15‰ to −17‰, i.e. around −3‰ lower than the δ¹³C measured for maize and faeces. Our study clearly demonstrates that the decomposition of complex substrates is associated with isotopic fractionation, causing evolved CO₂ to be depleted in ¹³C relative to substrates. Consequently the microbial products retained in the soil must be enriched in ¹³C.     

Three-source partitioning of CO2 efflux from soil planted with maize by C natural abundance fails due to inactive microbial biomass

A theoretical approach to the partitioning of carbon dioxide (CO₂) efflux from soil with a C₃ vegetation history planted with maize (Zea mays), a C₄ plant, into three sources, root respiration (RR), rhizomicrobial respiration (RMR), and microbial soil organic matter (SOM) decomposition (SOMD), was examined. The δ¹³C values of SOM, roots, microbial biomass, and total CO₂ efflux were measured during a 40-day growing period. A three-source isotopic mass balance based on the measured δ¹³C values and on assumptions made in other studies showed that RR, RMR, and SOMD amounted to 91%, 4%, and 5%, respectively. Two assumptions were thoroughly examined in a sensitivity analysis: the absence of ¹³C fractionation and the conformity of δ¹³C of microbial CO₂ and that of microbial biomass. This approach strongly overestimated RR and underestimated RMR and microbial SOMD. CO₂ efflux from unplanted soil was enriched in ¹³C by 2.0‰ compared to microbial biomass. The consideration of this ¹³C fractionation in the mass balance equation changed the proportions of RR and RMR by only 4% and did not affect SOMD. A calculated δ¹³C value of microbial CO₂ by a mass balance equation including active and inactive parts of microbial biomass was used to adjust a hypothetical below-ground CO₂ partitioning to the measured and literature data. The active microbial biomass in the rhizosphere amounted to 37% to achieve an appropriate ratio between RR and RMR compared to measured data. Therefore, the three-source partitioning approach failed due to a low active portion of microbial biomass, which is the main microbial CO₂ source controlling the δ¹³C value of total microbial biomass. Since fumigation-extraction reflects total microbial biomass, its δ¹³C value was unsuitable to predict δ¹³C of released microbial CO₂ after a C₃-C₄ vegetation change. The second adjustment to the CO₂ partitioning results in the literature showed that at least 71% of the active microbial biomass utilizing maize rhizodeposits would be necessary to achieve that proportion between RR and RMR observed by other approaches based on ¹⁴C labelling. The method for partitioning total below-ground CO₂ efflux into three sources using a natural ¹³C labelling technique failed due to the small proportion of active microbial biomass in the rhizosphere. This small active fraction led to a discrepancy between δ¹³C values of microbial biomass and of microbially respired CO₂.     

Use of 13C and 15N natural abundance techniques to characterize carbon and nitrogen dynamics in composting and in compost-amended soils

Isotope fractionation during composting may produce organic materials with a more homogenous δ¹³C and δ¹⁵N signature allowing study of their fate in soil. To verify this, C, N, δ¹³C and δ¹⁵N content were monitored during nine months covered (thermophilic; >40 °C) composting of corn silage (CSC). The C concentration reduced from 10.34 to 1.73 g C (g ash)⁻¹, or 83.3%, during composting. Nitrogen losses comprised 28.4% of initial N content. Compost δ¹³C values became slightly depleted and increasingly uniform (from −12.8±0.6‰ to −14.1±0.0‰) with composting. Compost δ¹⁵N values (0.3±1.3 to 8.2±0.4‰) increased with a similar reduced isotope variability.The fate of C and N of diverse composts in soil was subsequently examined. C, N, δ¹³C, δ¹⁵N content of whole soil (0-5 cm), light (<1.7 g cm⁻³) and heavy (>1.7 g cm⁻³) fraction, and (250-2000 μm; 53-250 μm and <53 μm) size separates, were characterized. Measurements took place one and two years following surface application of CSC, dairy manure compost (DMC), sewage sludge compost (SSLC), and liquid dairy manure (DM) to a temperate (C₃) grassland soil. The δ¹³C values and total C applied (Mg C ha⁻¹) were DM (−27.3‰; 2.9); DMC (−26.6‰; 10.0); SSLC (−25.9‰; 10.9) and CSC (−14.0‰; 4.6 and 9.2). The δ¹³C of un-amended soil exhibited low spatial (−28.0‰±0.2; n=96) and temporal (±0.1‰) variability. All C₄ (CSC) and C₃ (DMC; SSLC) composts, except C₃ manure (DM), significantly modified bulk soil δ¹³C and δ¹⁵N. Estimates of retention of compost C in soil by carbon balance were less sensitive than those calculated by C isotope techniques. One and two years after application, 95 and 89% (CSC), 75 and 63% (SSLC) and 88 and 42% (DMC) of applied compost C remained in the soil, with the majority (80-90%) found in particulate (>53 μm) and light fractions. However, C₄ compost (CSC) was readily detectable (12% of compost C remaining) in mineral (<53 μm) fractions. The δ¹⁵N-enriched N of compost supported interpretation of δ¹³C data. We can conclude that composts are highly recalcitrant with prolonged C storage in non-mineral soil fractions. The sensitivity of the natural abundance tracer technique to characterize their fate in soil improves during composting, as a more homogeneous C isotope signature develops, in addition to the relatively large amounts of stable C applied in composts.     

Effluxed CO2-C from sterilized and unsterilized treatments of a calcareous soil

Soil inorganic carbon (C) represents a substantial C pool in arid ecosystems, yet little data exist on the contribution of this pool to ecosystem C fluxes. A closed jar incubation study was carried out to test the hypothesis that CO₂-¹³C production and response to sterilization would differ in a calcareous (Mojave Desert) soil and a non-calcareous (Oklahoma Prairie) soil due to contributions of carbonate-derived CO₂. In addition to non-sterilized controls, soils were subjected to sterilization treatments (unbuffered HgCl₂ addition for Oklahoma soil and unbuffered HgCl₂ addition, buffered HgCl₂ addition, and autoclaving for Mojave Desert soil) to decrease biotic respiration and more readily measure abiotic CO₂ flux. Temperature and moisture treatments were also included with sterilization treatments in a factorial design.The rate of CO₂ production in both soils was significantly decreased (36-87%) by sterilization, but sterilization treatments differed in effectiveness. Sterilization had no significant effect on effluxed CO₂-¹³C values in the non-calcareous Oklahoma Prairie soil and autoclaved Mojave Desert soil as compared to their respective non-sterilized controls. However, sterilization significantly altered CO₂-¹³C values in Mojave Desert soil HgCl₂ sterilization treatments (both buffered and non-buffered). Plots of 1/CO₂ versus CO₂-δ¹³C (similar to Keeling plots) indicated that the source CO₂-δ¹³C value of the Oklahoma Prairie soil treatments was similar to the δ¹³C value of soil organic matter [(SOM); −17.76‰ VPDB] whereas the source for the (acidic) unbuffered-HgCl₂ sterilized Mojave Desert soil was similar to the δ¹³C value of carbonates (−0.93‰ VPDB). The source CO₂-δ¹³C value of non-sterilized and autoclaved (−18.4‰ VPDB) Mojave Desert soil treatments was intermediate between SOM (−21.43‰ VPDB) and carbonates and indicates up to 13% of total C efflux may be from abiotic sources in calcareous soils.     

Application of a quantum cascade laser-based spectrometer in a closed chamber system for real-time δC and δO measurements of soil-respired CO2

Laser spectroscopy is an emerging technique to analyze the stable isotopic composition of soil-respired CO₂ (δ¹³C_resp, δ¹⁸O_resp) in situ and at high temporal resolution. Here we present the first application of a quantum cascade laser-based spectrometer (QCLS) in a closed soil-chamber system to determine simultaneously δ¹³C_resp and δ¹⁸O_resp. In a Swiss beech forest, a total of 90 chamber measurements with 20 min sampling time each were performed. The instrument measured the δ¹³C and δ¹⁸O of the CO₂ in the chamber headspace at every second with a precision of 0.25‰, resulting in Keeling plots with 1200 data points. In addition, we calculated δ¹³C_resp directly from the flux ratio of ¹³CO₂ and ¹²CO₂. The flux-ratio values were 0.8‰ lower than the Keeling plot intercepts when the flux rates were derived from quadratic curve fits of the CO₂ increase. The δ¹⁸O-Keeling plots showed a significant bending very likely due to the equilibration of chamber CO₂ with the ¹⁸O of surface soil water. Therefore, we used a quadratic curve fit of the Keeling plots to estimate δ¹⁸O_resp. Our results also revealed that δ¹³C_resp was not constant throughout the CO₂ accumulation in the closed soil chambers: there were significant but non-systematic variations in δ¹³C_resp for the first 10 min, and systematic shifts in δ¹³C_resp of on average 1.9 ‰ in the second part of the 20-min measurements. These biases were probably caused by non-steady-state conditions in the soil-chamber system. Our study illustrates that the high temporal resolution of QCLS measurements allows the detection of non-linearities in the isotopic effluxes of CO₂ from the soil due to soil-chamber feedbacks. This information can be used to improve the estimates for δ¹³C_resp and δ¹⁸O_resp.     

Soil organic matter dynamics in grassland soils under elevated CO2: Insights from long-term incubations and stable isotopes

Elevated atmospheric carbon dioxide (CO₂) levels generally stimulate carbon (C) uptake by plants, but the fate of this additional C largely remains unknown. This uncertainty is due in part to the difficulty in detecting small changes in soil carbon pools. We conducted a series of long-term (170-330 days) laboratory incubation experiments to examine changes in soil organic matter pool sizes and turnover rates in soil collected from an open-top chamber (OTC) elevated CO₂ study in Colorado shortgrass steppe. We measured concentration and isotopic composition of respired CO₂ and applied a two-pool exponential decay model to estimate pool sizes and turnover rates of active and slow C pools. The active and slow C pools of surface soils (5-10 cm depth) were increased by elevated CO₂, but turnover rates of these pools were not consistently altered. These findings indicate a potential for C accumulation in near-surface soil C pools under elevated CO₂. Stable isotopes provided evidence that elevated CO₂ did not alter the decomposition rate of new C inputs. Temporal variations in measured δ¹³C of respired CO₂ during incubation probably resulted mainly from the decomposition of changing mixtures of fresh residue and older organic matter. Lignin decomposition may have contributed to declining δ¹³C values late in the experiments. Isotopic dynamics during decomposition should be taken into account when interpreting δ¹³C measurements of soil respiration. Our study provides new understanding of soil C dynamics under elevated CO₂ through the use of stable C isotope measurements during microbial organic matter mineralization.     

Three‐source partitioning of CO2 efflux from maize field soil by 13C natural abundance

A natural‐¹³C‐labeling approach—formerly observed under controlled conditions—was tested in the field to partition total soil CO₂ efflux into root respiration, rhizomicrobial respiration, and soil organic matter (SOM) decomposition. Different results were expected in the field due to different climate, site, and microbial properties in contrast to the laboratory. Within this isotopic method, maize was planted on soil with C₃‐vegetation history and the total CO₂ efflux from soil was subdivided by isotopic mass balance. The C₄‐derived C in soil microbial biomass was also determined. Additionally, in a root‐exclusion approach, root‐ and SOM‐derived CO₂ were determined by the total CO₂ effluxes from maize (Zea mays L.) and bare‐fallow plots. In both approaches, maize‐derived CO₂ contributed 22% to 35% to the total CO₂ efflux during the growth period, which was comparable to other field studies. In our laboratory study, this CO₂ fraction was tripled due to different climate, soil, and sampling conditions. In the natural‐¹³C‐labeling approach, rhizomicrobial respiration was low compared to other studies, which was related to a low amount of C₄‐derived microbial biomass. At the end of the growth period, however, 64% root respiration and 36% rhizomicrobial respiration in relation to total root‐derived CO₂ were calculated when considering high isotopic fractionations between SOM, microbial biomass, and CO₂. This relationship was closer to the 50% : 50% partitioning described in the literature than without fractionation (23% root respiration, 77% rhizomicrobial respiration). Fractionation processes of ¹³C must be taken into account when calculating CO₂ partitioning in soil. Both methods—natural ¹³C labeling and root exclusion—showed the same partitioning results when ¹³C isotopic fractionation during microbial respiration was considered and may therefore be used to separate plant‐ and SOM‐derived CO₂ sources.     

Plant biomass influences rhizosphere priming effects on soil organic matter decomposition in two differently managed soils

We used a continuous labeling method of naturally ¹³C-depleted CO₂ in a growth chamber to test for rhizosphere effects on soil organic matter (SOM) decomposition. Two C3 plant species, soybean (Glycine max) and sunflower (Helianthus annus), were grown in two previously differently managed soils, an organically farmed soil and a soil from an annual grassland. We maintained a constant atmospheric CO₂ concentration at 400±5 ppm and δ¹³C signature at −24.4‰ by regulating the flow of naturally ¹³C-depleted CO₂ and CO₂-free air into the growth chamber, which allowed us to separate new plant-derived CO₂-C from original soil-derived CO₂-C in soil respiration. Rhizosphere priming effects on SOM decomposition, i.e., differences in soil-derived CO₂-C between planted and non-planted treatments, were significantly different between the two soils, but not between the two plant species. Soil-derived CO₂-C efflux in the organically farmed soil increased up to 61% compared to the no-plant control, while the annual grassland soil showed a negligible increase (up to 5% increase), despite an overall larger efflux of soil-derived CO₂-C and total soil C content. Differences in rhizosphere priming effects on SOM decomposition between the two soils could be largely explained by differences in plant biomass, and in particular leaf biomass, explaining 49% and 74% of the variation in primed soil C among soils and plant species, respectively. Nitrogen uptake rates by soybean and sunflower was relatively high compared to soil C respiration and associated N mineralization, while inorganic N pools were significantly depleted in the organic farm soil by the end of the experiment. Despite relatively large increases in SOM decomposition caused by rhizosphere effects in the organic farm soil, the fast-growing soybean and sunflower plants gained little extra N from the increase in SOM decomposition caused by rhizosphere effects. We conclude that rhizosphere priming effects of annual plants on SOM decomposition are largely driven by plant biomass, especially in soils of high fertility that can sustain high plant productivity.     

Experiments on the origin of C in the calcium carbonate granules produced by the earthworm Lumbricus terrestris

Granules of calcium carbonate up to 2.5 mm in size have been found in archaeological soils and sediments for many years. Recent work has shown conclusively that these granules are derived from the calciferous glands of earthworms and most commonly (in Northern European soils) from Lumbricus terrestris. Valuable pedological and archaeological information could be obtained from a radiocarbon date if the ¹⁴C in the granule carbonate reflects the true age of formation. Preliminary information on the possible sources of carbon in the granule calcite was obtained by a series of simple experiments growing worms in artificial substrates made up of materials with marked differences in δ¹³C. The results showed that carbon in the granule calcite comes from the dietary inputs, but may be coming partly from atmospheric CO₂ and also partly from old soil carbon. Little appears to come from the soil calcium carbonate. Most of the results imply that a fractionation of around +12‰ is occurring between the carbon ingested by the worm and the carbon output in the granules. This is comparable to (but slightly higher than) the widely quoted non-biological value (ca. +10‰) of total kinetic fractionation between CO₂-CaCO₃.     

Isotopic analysis of respired CO2 during decomposition of separated soil organic matter pools

A detailed understanding of the processes that contribute to the δ¹³C value of respired CO₂ is necessary to make links between the isotopic signature of CO₂ efflux from the soil surface and various sources within the soil profile. We used density fractionation to divide soils from two forested sites that are a part of an ongoing detrital manipulation experiment (the Detrital Input and Removal Treatments, or DIRT project) into two soil organic matter pools, each of which contributes differently to total soil CO₂ efflux. In both sites, distinct biological pools resulted from density fractionation; however, our results do not always support the concept that the light fraction is readily decomposable whereas the heavy fraction is recalcitrant. In a laboratory incubation following density fractionation we found that cumulative respiration over the course of the incubation period was greater from the light fraction than from the heavy fraction for the deciduous site, while the opposite was true for the coniferous site.Use of stable isotopes yielded insight as to the nature of the density fractions, with the heavy fraction solids from both forests isotopically enriched relative to those of the light fraction. The isotopic signature of respired CO₂, however, was more complicated. During incubation of the fractions there was an initial isotopic depletion of the respired CO₂ compared to the substrate for both soil fractions from both forests. Over time for both fractions of both soils the respired δ¹³C reflected more closely the initial substrate value; however, the transition from depleted to enriched respiration relative to substrate occurs at a different stage of decomposition depending on site and substrate recalcitrance. We found a relationship between cumulative respiration during the incubation period and the duration of the transition from isotopically depleted to enriched respiration in the coniferous site but not the deciduous site. Our results suggest that a shift in microbial community or to dead microbial biomass as a substrate could be responsible for the transition in the isotopic signature of respired CO₂ during decomposition. It is likely that a combination of organic matter quality and isotopic discrimination by microbes, in addition to differences in microbial community composition, contribute to the isotopic signature of different organic matter fractions. It is apparent that respired δ¹³CO₂ cannot be assumed to be a direct representation of the substrate δ¹³C. Detailed knowledge of the soil characteristics at a particular site is necessary to interpret relationships between the isotopic values of a substrate and respired CO₂.     

Microbial use of maize cellulose and sugarcane sucrose monitored by changes in the C/C ratio

An arable soil with organic matter formed from C₃-vegetation was amended initially with maize cellulose (C₄-cellulose) and sugarcane sucrose (C₄-sucrose) in a 67-day laboratory incubation experiment with microcosms at 25 °C. The amount and isotopic composition (¹³C/¹²C) of soil organic C, CO₂ evolved, microbial biomass C, and microbial residue C were determined to prove whether the formation of microbial residues depends on the quality of the added C source adjusted with NH₄NO₃ to the same C/N ratio of 15. In a subsequent step, C₃-cellulose (3 mg C g⁻¹ soil) was added without N to soil to determine whether the microbial residues formed initially from C₄-substrate are preferentially decomposed to maintain the N-demand of the soil microbial community. At the end of the experiment, 23% of the two C₄-substrates added was left in the soil, while 3% and 4% of the added C₄-cellulose and C₄-sucrose, respectively, were found in the microbial biomass. The addition of the two C₄-substrates caused a significant 100% increase in C₃-derived CO₂ evolution during the 5-33 day incubation period. The addition of C₃-cellulose caused a significant 50% increase in C₄-derived CO₂ evolution during the 38-67 day incubation period. The decrease in microbial biomass C₄-C accounted for roughly 60% of this increase. Cellulose addition promoted microorganisms strongly able to recycle N immediately from their own tissue by “cryptic growth” instead of incorporating NO₃⁻ from the soil solution. The differences in quality of the microbial residues produced by C₄-cellulose and C₄-sucrose decomposing microorganisms are also reflected by the difference in the rates of CO₂ evolution, but not in the rates of net N mineralization.     

Dynamics of maize (Zea mays L.) leaf straw mineralization as affected by the presence of soil and the availability of nitrogen

An incubation experiment was carried out with maize (Zea mays L.) leaf straw to analyze the effects of mixing the residues with soil and N amendment on the decomposition process. In order to distinguish between soil effects and nitrogen effects for both the phyllospheric microorganisms already present on the surface of maize straw and soil microorganisms the N amendment was applied in two different placements: directly to the straw or to the soil. The experiment was performed in dynamic, automated microcosms for 22 days at 15 °C with 7 treatments: (1) untreated soil, (2) non-amended maize leaf straw without soil, (3) N amended maize leaf straw without soil, (4) soil mixed with maize leaf straw, (5) N amended soil, (6) N amended soil mixed with maize leaf straw, and (7) soil mixed with N amended maize leaf straw. ¹⁵NH₄¹⁵NO₃ (5 at%) was added. Gas emissions (CO₂, ¹³CO₂ and N₂O) were continuously recorded throughout the experiment. Microbial biomass C, biomass N, ergosterol, δ¹³C of soil organic C and of microbial biomass C as well as ¹⁵N in soil total N, mineral N and microbial biomass N were determined in soil samples at the end of the incubation. The CO₂ evolution rate showed a lag-phase of two days in the non-amended maize leaf straw treatment without soil, which was completely eliminated when mineral N was added. The addition of N generally increased the CO₂ evolution rate during the initial stages of maize leaf straw decomposition, but not the cumulative CO₂ production. The presence of soil caused roughly a 50% increase in cumulative CO₂ production within 22 days in the maize straw treatments due to a slower decrease of CO₂ evolution after the initial activity peak. Since there are no limitations of water or N, we suggest that soil provides a microbial community ensuring an effective succession of straw decomposing microorganisms. In the treatments where maize and soil was mixed, 75% of microbial biomass C was derived from maize. We concluded that this high contribution of maize using microbiota indicates a strong influence of organisms of phyllospheric origin to the microbial community in the soil after plant residues enter the soil.     

Annual variation in δC values of maize and wheat: Effect on estimates of decadal scale soil carbon turnover

On sites where C4-plants have replaced C3-plants, changes in soil δ¹³C allow the turnover of C3- and C4-derived C to be separated. Studies of decadal scale turnover of soil C following conversion to C4-plants generally lack δ¹³C values for previous C4-residue inputs and assume that estimates of C4-derived soil C to be based on a fixed δ¹³C value. Further assumptions are that changes in the initial (time-zero) soil δ¹³C values are insignificant following conversion to C4-plants. We tested these assumptions by measuring: 1) the δ¹³C of annual samples of silage maize biomass (C4-plant) and winter wheat grains (C3-plant) grown during 1988 to 2006, and 2) the δ¹³C of soil kept under bare fallow during 1956 to 1983. The δ¹³C of plants was related to climate variables, and the impact of maize δ¹³C was based on estimates of maize-derived soil C using different approaches to establish the δ¹³C in maize inputs. The δ¹³C of both maize and wheat decreased with time, but the rate of change and annual variations were considerably larger for wheat than for maize. Maize as well as wheat δ¹³C was best related to year (probably reflecting a decrease in atmospheric δ¹³C) and the water balance during the active growth period. Using the smallest (−12.44‰) and the largest (−11.26‰) δ¹³C measured during 1988 and 2006, estimates of maize-derived C in soil after 18 years ranged from 13.2% to 14.2% of the soil total C. Despite a loss of 31% of the soil C pool under bare fallow, the increase in soil δ¹³C was significant only at P < 0.10. We conclude that annual variations in maize δ¹³C values and changes in the δ¹³C of the soil C fraction derived from the pre-conversion C3-vegetation have only little impact on estimates of maize-derived soil C that cover a few decades. For estimates covering several decades to centuries, the subtle but consistent changes in plant and soil δ¹³C need to be accounted for. The variability in δ¹³C in wheat grains suggest that the use of any fixed δ¹³C value for C3-residues in estimates of C turnover in soils on which C4-plants have been replaced by C3-plants can be associated with considerable uncertainty.     

Organic carbon and stable C isotope in conservation agriculture and conventional systems

Conservation agriculture might have the potential to increase soil organic C content compared to conventional tillage based systems. The present study quantified soil organic carbon (SOC) and soil C derived from C₃ (wheat) and C₄ (maize) plant species using δ¹³C stable isotope. Soil with 16 y of continuous application of zero tillage (ZT) or conventional tillage (CT), monoculture (M) or rotation (R) of wheat and maize, and with (+r) and without retention (−r) in the field of crop residues were studied in the central highlands of Mexico. The highest SOC content was found in the 0-5 cm layer under ZTM and ZTR with residues retention. The soil cultivated with maize showed a higher SOC content in the 0-10 cm layer with residue retention than without residue. In the 10-20 cm layer, the highest SOC content was found in the CT treatment with residue retention. The SOC stock expressed as equivalent soil mass was greatest in the 0-20 cm layer of the ZTM (wheat and maize) and ZTR cultivated treatments with residue retention. After 16 y, the highest content of soil δ¹³C was found in ZTM + r and CTM + r treated soil cultivated with maize; −16.56‰ and −18.08‰ in the 0-5 cm layer, −18.41‰ and −18.02‰ in the 5-10 cm layer and −18.59‰ and −18.72‰ in the 10-20 cm layer respectively. All treatments had a higher percentages of C-C₃ (derived from wheat residues or the earlier forest) than C-C₄ (derived from maize residues). The highest percentages of C-C₄, was found in ZTM + r and CTM + r treated soil cultivated with maize, i.e. 33.0% and 13.0% in 0-5 cm layer, 9.1% and 14.3% in the 5-10 cm layer and 5.0% and 6.8% in 10-20 cm layer, respectively. The gross SOC turnover was lower in soil with residue retention than without residues. It was found that the ZT system with residue retention and rotation with wheat is a practice with a potential to retain organic carbon in soil.     

Use of C abundance to study short-term pig slurry decomposition in the field

Applying pig slurry (PS) on agricultural soils is a common practice. However, its impact on soil organic C dynamics is not clear. This experiment investigated the use of natural ¹³C abundance to study the short-term C mineralization of anaerobically stored PS under field conditions. Measurements of δ¹³C-CO₂ were made on soil air samples obtained from a bare sandy loam during 22 d following incorporation of either PS alone, PS+barley straw, or barley straw alone; an unamended treatment was used as a control. Slurry C was enriched in ¹³C (−20.0‰) because of the high corn (Zea mays L.) content of the animal diet. This value contrasted with δ¹³C of −28.4‰ for the soil organic matter and of −29.0‰ for the barley straw. A peak of high δ¹³CO₂ values (average of −9.2‰) was observed on the day of PS application and was attributed to the dissociation of PS carbonates when mixed with the relatively acidic soil. After this initial burst, 36% of the evolved CO₂ originated from the decomposing PS. After 22 d of incubation, approx. 20% of the PS-C had been lost as CO₂. This short-term field study did not show any priming effect of PS on the mineralization of straw or native soil C. Due to its heterogeneity, the use of the isotopic composition of the evolved CO₂ for estimating PS decomposition requires precaution either through the use of a specific experimental design involving comparable C3 and C4 treatments, or calculations to account for the presence of ¹³C-enriched inorganic C in the PS.     

Abiotic drivers and their interactive effect on the flux and carbon isotope (C and δC) composition of peat-respired CO2

Feedbacks to global warming may cause terrestrial ecosystems to add to anthropogenic CO₂ emissions, thus exacerbating climate change. The contribution that soil respiration makes to these terrestrial emissions, particularly from carbon-rich soils such as peatlands, is of significant importance and its response to changing climatic conditions is of considerable debate. We collected intact soil cores from an upland blanket bog situated within the northern Pennines, England, UK and investigated the individual and interactive effects of three primary controls on soil organic matter decomposition: (i) temperature (5, 10 and 15 °C); (ii) moisture (50 and 100% field capacity – FC); and (iii) substrate quality, using increasing depth from the surface (0–10, 10–20 and 20–30 cm) as an analogue for increased recalcitrance of soil organic material. Statistical analysis of the results showed that temperature, moisture and substrate quality all significantly affected rates of peat decomposition. Q₁₀ values indicated that the temperature sensitivity of older/more recalcitrant soil organic matter significantly increased (relative to more labile peat) under reduced soil moisture (50% FC) conditions, but not under 100% FC, suggesting that soil microorganisms decomposing the more recalcitrant soil material preferred more aerated conditions. Radiocarbon analyses revealed that soil decomposers were able to respire older, more recalcitrant soil organic matter and that the source of the material (deduced from the δ¹³C analyses) subject to decomposition, changed depending on depth in the peat profile.     

Microbial response to exudates in the rhizosphere of young beech trees (Fagus sylvatica L.) after dormancy

Plants act as an important link between atmosphere and soil: CO2 is transformed into carbohydrates by photosynthesis. These assimilates are distributed within the plant and translocated via roots into the rhizosphere and soil microorganisms. In this study, 3 year old European beech trees (Fagus sylvatica L.) were exposed after the chilling period to an enriched ¹³C–CO₂ atmosphere (δ¹³C = 60‰ – 80‰) at the time point when leaves development started. Temporal dynamics of assimilated carbon distribution in different plant parts, as well as into dissolved organic carbon and microbial communities in the rhizosphere and bulk soil have been investigated for a 20 days period. Photosynthetically fixed carbon could be traced into plant tissue, dissolved organic carbon and total microbial biomass, where it was utilized by different microbial communities. Due to carbon allocation into the rhizosphere, nutrient stress decreased; exudates were preferentially used by Gram-negative bacteria and (mycorrhizal) fungi, resulting in an enhanced growth. Other microorganisms, like Gram-positive bacteria and mainly micro eucaryotes benefited from the exudates via food web development. Overall our results indicate a fast turnover of exudates and the development of initial food web structures. Additionally a transport of assimilated carbon into bulk soil by (mycrorhizal) fungi was observed.     

Microbial immobilisation of C rhizodeposits in rhizosphere and root-free soil under continuous C labelling of oats

A greenhouse experiment was conducted by growing oats (Avenasativa L.) in a continuously ¹³CO₂ labeled atmosphere. The allocation of ¹³C-labeled photosynthates in plants, microbial biomass in rhizosphere and root-free soil, pools of soil organic C, and CO₂ emissions were examined over the plant's life cycle. To isolate rhizosphere from root-free soil, plant seedlings were placed into bags made of nylon monofilament screen tissue (16 μm mesh) filled with soil. Two peaks of ¹³C in rhizosphere pools of microbial biomass and dissolved organic carbon (DOC), as well as in CO₂ emissions at the earing and ripeness stages were revealed. These ¹³C maxima corresponded to: (i) the end of rapid root growth and (ii) beginning of root decomposition, respectively. The δ¹³C values of microbial biomass were higher than those of DOC and of soil organic matter (SOM). The microbial biomass C accounted for up to 56 and 39% of ¹³C recovered in the rhizosphere and root-free soil, respectively. Between 4 and 28% of ¹³C assimilated was recovered in the root-free soil. Depending on the phenological stage, the contribution of root-derived C to total CO₂ emission from soil varied from 61 to 92% of total CO₂ evolved, including 4-23% attributed to rhizomicrobial respiration. While 81-91% of C substrates used for microbial growth in the root-free soil and rhizosphere came from SOM, the remaining 9-19% of C substrates utilized by the microbial biomass was attributable to rhizodeposition. The use of continuous isotopic labelling and physical separation of root-free and rhizosphere soil, combined with natural ¹³C abundance were effective in gaining new insight on soil and rhizosphere C-cycling.